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Revision as of 02:04, 14 October 2018
Notebook-molecular biology
18.07.18
PPK transformation
18.08.05
Obtained data on changes in phosphorus concentration
18.08.27
Get toxin antitoxin 2 DNA
18.08.28
Complete ppx+ppn pathway release phosphorus detection
18.09.01
Sequence amplification
18.09.02
Not-gate fusion into the light control promoter
18.09.06
The plasmid that ultimately obtains the not-gate sequence fusion into the light control promoter
18.09.07
Light induced pathway construction
18.09.03
The target fragment of toxin antitoxin 2 is linked to psb1c3 backbone
18.09.07
first round verification of toxin antitoxin 2
18.09.09
All ligation products are transformed and transformed into chloramphenicol resistant plates(light control pathway)
18.09.10
Identification and expansion of single colonies produced by transformation(light control pathway)
18.09.11
Verification of the function of the light control pathway
18.09.11
second round verification of toxin antitoxin 2
18.09.12
In vitro expression verification protein function(light control pathway)
18.09.16
Third round verification of toxin antitoxin2
18.09.20
Complete PPN protein expression detection
18.09.25
Get toxin antitoxin1, dCBD-OmpA, Lectin-OmpA, FimH-lectin DNA
18.09.29
First round verification of toxin antitoxin1
18.10.01
Phosphorus release effect of ppn, ppx was detected by Albert staining
18.10.01
Completed the design and assembly of ppx+rbs part
18.10.01
The target fragment of toxin antitoxin 1, dCBD-OmpA, Lectin-OmpA, FimH-lectin are linked to psb1c3 backbone
Below is part of our raw data
Theophylline concentrationg/L |
0 |
0.5 |
1 |
2 |
4 |
6 |
8 |
OD600 6h |
0.397 |
0.174 |
0.182 |
0.075 |
0.086 |
0.074 |
0.066 |
0.358 |
0.214 |
0.166 |
0.08 |
0.07 |
0.067 |
0.08 |
|
0.305 |
0.242 |
0.1 |
0.088 |
0.082 |
0.065 |
0.095 |
|
0.353333 |
0.21 |
0.149333 |
0.081 |
0.079333 |
0.068667 |
0.080333 |
|
OD600 12h |
0.547 |
0.29 |
0.264 |
0.286 |
0.101 |
0.064 |
0.074 |
0.46 |
0.353 |
0.24 |
0.15 |
0.239 |
0.085 |
0.083 |
|
0.575 |
0.271 |
0.212 |
0.115 |
0.143 |
0.106 |
0.09 |
|
0.527333 |
0.304667 |
0.238667 |
0.183667 |
0.161 |
0.085 |
0.082333 |
|
0 |
0.1 |
0.2 |
0.4 |
0.8 |
1.6 |
||
OD600 8H |
0.37 |
0.415 |
0.407 |
0.347 |
0.319 |
0.131 |
|
0.46 |
0.374 |
0.438 |
0.303 |
0.321 |
0.172 |
||
0.41 |
0.461 |
0.456 |
0.42 |
0.302 |
0.153 |
||
0.413333 |
0.416667 |
0.433667 |
0.356667 |
0.314 |
0.152 |
||
OD600 16H |
0.691 |
0.704 |
0.732 |
0.613 |
0.573 |
0.35 |
|
0.704 |
0.725 |
0.714 |
0.738 |
0.561 |
0.464 |
||
0.732 |
0.697 |
0.748 |
0.646 |
0.581 |
0.442 |
||
0.709 |
0.708667 |
0.731333 |
0.665667 |
0.571667 |
0.418667 |
IPTG induction time |
ccas+Not Gate（induced） |
ccas+GFP（induced） |
ccas+ccar empty cotrol |
positive control |
medium |
|
0h |
11649.5 |
11380 |
11468.25 |
12249 |
10811.5 |
|
1h |
11969.5 |
11744.5 |
11811.75 |
14048.5 |
||
2.5h |
13396.75 |
13640.5 |
13005 |
18427.5 |
||
5h |
15668.5 |
15552 |
12744.75 |
20083.75 |
||
|
||||||
0h |
838 |
568.5 |
656.75 |
1437.5 |
||
1h |
1158 |
933 |
1000.25 |
3237 |
||
2.5h |
2585.25 |
2829 |
2193.5 |
7616 |
||
5h |
4857 |
4740.5 |
1933.25 |
9272.25 |
||
with or without light for 2h |
||||||
green light for 2hs |
14614 |
18971 |
13837.25 |
20295.75 |
||
red light for 2hs |
16023.5 |
16166.75 |
14467 |
20026 |
||
dark for 2hs |
15905.75 |
15803 |
13965.25 |
19268.75 |
||
|
||||||
green light for 2hs |
3802.5 |
8159.5 |
3025.75 |
9484.25 |
||
red light for 2hs |
5212 |
5355.25 |
3655.5 |
9214.5 |
||
dark for 2hs |
5094.25 |
4991.5 |
3153.75 |
8457.25 |
||
continue to culture for 1h (Green to Red, Red to Green） |
||||||
Cultivate with green light for 2 hrs and then cultivating with red light for 1hr |
15819.25 |
17099.5 |
14089.83333 |
19863.5 |
||
Cultivate with red light for 2hrs and then cultivating with green light for 1hr |
15099.75 |
17902 |
14216.125 |
19647.375 |
||
dark for 3hs |
16065 |
16051.75 |
13965.25 |
19268.75 |
||